Chemotherapeutic methods and compositions
Disclosed herein are methods and compositions for enhancing the cell-killing activity of anti-neoplastic agents by inhibiting the activity of a lysyl oxidase-type enzyme. Also disclosed are methods for screening for chemotherapeutic agents, and for molecules that enhance the activity of chemotherapeutic agents, using cells grown on an extracellular matrix.
1. A method for killing a malignant cell, the method comprising: contacting the malignant cell, wherein the malignant cell expresses a lysyl oxidase-type enzyme, with an anti-neoplastic agent and an inhibitor of the lysyl oxidase-type enzyme, wherein the cell is growing in the presence of an extracellular matrix containing collagen, whereby the malignant cell is killed, wherein the anti-neoplastic agent is a chemotherapeutic agent and the inhibitor is an isolated lysyl oxidase-like 2 (LOXL2) antibody or antigen-binding fragment thereof specifically binds to an epitope within the amino acid sequence of SEQ ID NO: 28; wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 15.
2. The method of claim herein the malignant cell is in culture.
3. The method of claim 1 , wherein the malignant cell is present in a tumor.
4. The method of claim 1 , wherein the chemotherapeutic agent is selected from the group consisting of methotrexate, cispiatin, doxorubicin, docetaxel, erlotinih, paciitaxel, paraclitaxel, 5-fluorouracil and germcitibine.
5. The method of claim 1 , wherein the collagen comprises type 1 collagen.
6. The method of claim 1 , wherein the LOXL2 antibody or antigen-binding fragment thereof is a Fv, a scFv, a Fab, or a F(ab′) 2 .
7. The method of claim 1 , wherein the LOXL2 inhibitor inhibits LOXL2 enzymatic activity.
8. The method of claim 1 , wherein the LOXL2 antibody or antigen-binding fragment thereof is polyclonal.
9. The method of claim 1 , wherein the antibody or antigen-binding fragment thereof is monoclonal.
10. The method of claim 9 , wherein the antibody or antigen-binding fragment thereof is humanized.
11. The method of claim 1 , wherein the antibody or antigen-binding fragment thereof comprises the complementarity determining regions CDRs), CDR1, CDR2, and CDR3, of a heavy chain variable region comprising the amino acid sequence of SEQ ID NO; 14, and the CDRs, CDR1, CDR2, and CDR3, of a light chain variable region comprising the amino acid sequence of SEQ ID NO: 15.
12. The method of claim 11 , wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEC) ID NO: 14.
13. The method of claim 11 , wherein the antibody or antigen-binding fragment thereof comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO; 15.
14. The method of claim 1 , wherein the malignant cell is selected from the group consisting of fibrosarcoma, breast carcinoma, breast adenocarcinoma, pancreatic adenocarcinoma, pancreatic tumor, colon adenocarcinoma, ovarian adenocarcinoma, and lung carcinoma.